CA1118768A

CA1118768A - Peptides

Info

Publication number: CA1118768A
Application number: CA000310850A
Authority: CA
Inventors: Janos Pless; Edmond Sandrin
Original assignee: Sandoz AG
Current assignee: Sandoz AG
Priority date: 1977-09-07
Filing date: 1978-09-07
Publication date: 1982-02-23
Also published as: ATA639878A; EP0001061A1; IE781798L; IT1099493B; DK381978A; US4187295A; ES473071A1; PH14006A; IE47341B1; NZ188345A; EP0001061B1; JPS5448758A; DE2860579D1; FI64798C; IL55507A; FI782641A; FI64798B; PT68522A; IT7827432A0; AT372075B

Abstract

Abstract of the Dlsclosure The invention provides polypeptldes useful, for example, as soporifics, a process for the preparation thereof and com,oositions containing these compounds. They have the formula A - B - C - D - E
wherein A is H-Trp, H-MeTrp, 9-Trp(5-OH) or H-MeTrp (5-OH) B is Ala C is Ser, Thr, Ala, Gly, Val or a residue of formula

Description

100-48~1 IMPROV~MENTS IN OR RELATING TO ORGANIC COMPOUNDS
. _ _ _ _ ~The present invention relates to polypeptide derivatives, More particularly, the present invention provides compounds of formula I, A - B - C - D - E T

- wherein A is H-Trp, H-MeTrp, H-Trp(500H~ or . H-MeTrp(5-OH), B is Ala, C i5 Ser, Thr~ Ala, Gly, Val, a restdue of formula .
-NH-CH-CO- or -NH-CH-CO- OR

- wherein R is hydrogen, alkyl of 1 to 4 carbon atoms, or an alkali metal or an ~3 `
"/

- 2 - 100-4881 alkali.ne e~rth metal and Rl is hydrogen or methyl, D is Gly or Sar, is a residue -N~-CH-CH2-Y, , X
wherein X and Y independently signify COZ or CH20H/
z is OH, OR2, NH2, NHR3 or NR3R4~ a~d each of R2, R3 and R~ independently signifies alkyl of 1 to 4 car~on atoms whereby the residues A, B, C and.~ can have the L , D- or D, L conigeration.
The residue A can, for example, be H-Trp or H ~eTrp. Additionally, A can be H-Trp(5-OH) or H-MeTrp(5-OH).
The residue C can be Ser, Thr, Ala, Gly or Val. Additionally, C can be a residue -NH-CN-CO-. - R
wherein R and Rl are as previously deined.~ C can also be a res~due -NH-CH-CO- OR
CH-O-P /
R ~ \OR
wherein ]R and Rl are as previously defined.

'. ! ! ' ',: , , ' ' , .
: .

L87~3 ~ 3 - 100-4~81 In one group or compounds R is hydrogen or alkyl of 1 to 4 carbon atoms. In another group of compounds R is an alkali or al~aline earth metal.
The residue E can be a group -NH-CHX-CH2-Y
wherein X and Y are as previously de~ined. When X andt or Y is/are COZ, Z can be OH or OR2 whereln R2 is as previously defined. Z can also be NH2, NHR3 or NR3R4 ~herein R3 and ~ are as previously defined.
A peptide of formula I can be obtained by methods which are conventional in the art.
Accordingly, the present invention provides a process for the production o~ a peptide of formula I
which comprises, a~ removing at least one protectlve group from a protec~ed peptide having the sequence indicated in formula I, or b~ lin~ing together by an amide bond two peptide units each of which contains at least one amino acid and which is in protected or unprotected form, the peptide 2~ units being such that thP amino acid sequence given in formula I is obtained, ~nd then, if necessary, effect-ing process variant a~, or c) converting a group E of an unprotected or protected peptide into another group E having the definition :

~Li876~

previously indicated, and, if necessary effecting process variant a)~
The above methods are known in peptide chemistry and may be effected in manner analogous to the processes described in the following Examples.
Insofar as the production of the starting materials is not particularly described, these compounds are known or may be produced and purified in accordance with known methods. These compounds may also be produced in a manner analogous to the procPsses described in the following Examples.
The compounds may exlst in salt form, in-cluding acid addition salt forms or in the form of complexes, for example, complexes with metals. Suitable acids for acid addition salt formation include organic acids, polymeric acids and lnorganic acids. Complexes may, or example, be formed with elements such as metals, e.g. calcium, magnesium, aluminium, cobalt and especially zinc.
In the following Examples, all temperatures are indicated in degrees Celsius.
The following abbreviations are used:
Asn = asparagine residue Asn-ol = asparaginol residue Asp-diol- asparagin~diol-= 2-amino-butane-1,4-6~3 - 5 - . 100-4881 = diol residue Asp(~HCH3~-ol = N'-methylasparaginol residue Asp(OMe)~OMe = aspartic acid dimethyl est~r residue DMF ~ dimethyl formamide Et = ethyl Me = methyl kle~rp = N-methyltryptophan residue (5-OH)-Trp - 5-hydroxytryptophan residue OTcp = 2,4,5-trichlorophenoxy TFA = trifluoroacetic acid.
Z = benzyloxycarbonyl All of the amino acid residues with the exceptlon of glycyl, as ~ell as all of the amino alcohol residues referred to in this specification, possess the L- configuration unless other~ise stated.
An amino alcohol ~s associated wlth the L- series when the CH2OH group is in the same position as the ~ -COOH group in the corresponding L amino acid.

3761~3 .

EYAMPLE 1: H-Trp-Ala-Ser-Gly-L-Asn--ol To a solution of 2.0 g of Z-Trp-Ala-Ser-Gly-L-Asn-ol in 80 ml of dioxanle and 3.2 ml of aqueous IN HCL is added 1 g of a palladium catalyst and hydrogenation is effected at room temperature and normal pressure until no further hydrogan is taken up. The solution is filtered, the solvent evaporated and the residue ls trlturated with ether. The resulting title compound in the form of the hydrochloride salt, decom-poses~at 170, [o~]20 = -15.3 ~C= 1.0 ln water).
The Z-Trp-Ala--Ser-Gly-L-Asn-ol used as starting material is prepared as follows: - -a) Z-Trp-Ala-Ser-Gly-NHNH2 _________ _____________ 10.5 g of 2-Trp-OTcp are added to a solution o~ 6.0 g of H-Ala-Ser-Gly-OEt. hydrochloride and 2.8 ml of triethylamine in 50 ml of DME. After standing for 24 hours at room temperature the solvent ~s removed in vacuo and the residue ~riturated with dilute aqueous - HCl and ethyl acetate. The residue is dissolved in 80 ml of DMF and 12 ml hydrazine hydrate are added. After standing for 24'hours the title compound is precipi-tated by adding ether. The precipitate is filtered, washed with sthanol and drie~.
b) Z-Tr~-Ala-Ser~Gl~-L-Asn-ol ____ ___________ _________ 2.9 ml of 5.6~ HCl ln dioxan and 0.67 ml of 8~ ~ ~

~ 7 ~ 100 tert. butyl nitrite are added at -~0 to a solution of

3,2 g of Z-Trp-Ala-Ser-Gly-NHNH2 in 50 ml of DMF. After standing 15 minutes at -20 1.89 of L-Asparaginol hydrochloride and 2.5 ml of triethylamine are added.
After standing for 15 hours at room temperature the reaction mlxture is considerably reduced ln volume, diluted witn ethyl acetata and washed with dilute aqueous hydrochloric acid and with water. The title compound precipitates from the concentrated solution.

The following compounds of formula I (wherein D = Gly) can be prepared in manner analogous to that o~ Example 1 using appropriate starting materials in approxlmately equlvalent amounts.

376~
- 8 - 100~B81 _~- x. - _ B C ~ Salt- -[a32 . . . , ,, . __ _ __ _ __ 2 H-Trp Ala Ser Asp-OH HCl ~ l,Oa 3 H-Trp Ala Ser Asn-OH ~Cl -16,2

4 H-Trp Ala Ser Asn-OMe HCl -17,0b .
H-Trp Ala Ser Asp-diol HCl ~20,0~ .
6 H-Trp D-Ala Ser A~n-ol HCl ~14,9a .
7 H-Trp D-Ala Ser Asp-OH HCl ~ 4, 8 H-Trp D-Ala Ser Asn-OH HCl ~ 8.7a 9 ~-Trp D-~la Ser Asn-OMe HCl ~ 6,3a .-H-Trp D-Ala Ser Asp-diol HCl ~ 7,9a 11 H-D-Trp Ala Ser Asn-ol HCl -86,3b 12 H-D-Trp Ala Ser Asp-OH HCl _75,7b 13 H-D-Trp D-Ala Ser Asn-ol ~Cl _72,~b 14 H-D-Trp D-Ala Ser Asp-OH HCl -10 b H-MeTrp Ala Ser Asn-ol TFA -10,3b 16 H-Me~rp Ala ~er ~sp-OH TFA -13,5b .
17 H-Trp Ala Gly Asp-OH HCl ~17,6a 18 H-Trp D-Ala Thr Asn-ol HCl ~18,7a 19 H-Trp D-Ala Thr Asp-OH HCl ~20,9 2~ H-Trp D-Ala Ser Asp(OMe)-OMe HCl ~ 2,4a 21H- ~5-OH) -Trp D-Ala 5er Asp-OH TFA ~27,7 22 H-Trp . . ~la V~l Asn-ol HCl l~7a 23 H-Trp D Al~ ~ly Asp-OH HCl +29,8C
24 H-Trp D-Ala Ser ~sp(NHCH3)-ol HCl ~18,0d H-Trp D-Ala Val Asp-diol HCl ~16,7e . . ~ 1 ----- ~ ..
a: c = 1,0 in`95~ acetic acid. ~ .
b: c = 1,0 in water c: c = 0,55 in 95% acetic acid d: c = 0,51 in 95~ acetic acid e: c = 0.96 in 95% acetic acld - 9 - 100-4~81 The compounds of formula I exhlbit pharma-cological activity. In particular, the compounds exhibit central nervous system activity as indicated in standard tests in anima:Ls r for example as indicated by the method ~P.O.T.) described in S. Irwin, Gordon Research Conference, Medic.Lnal Chemistry, 1959 r J.~. Nodide and P.E. Siegler, Animal and Clinical Pharmacologic Techniques in Drug Evaluation, Chicago 1364 and in Psychopharmacologia 13, 222-257, 1963 (Berlin~. The indlcated activity, can either suppress - or stimulate the central nervous system, depending on the structure and dosage of the compound administered.
These activities can also be demonstrated in mice by means of the Motron test. In this test, the motor activity of the animals is determined electroni-cally using a motillty measurlng apparatus. For each dose, two groups each comprising ive mice (one group as a control group) are set up and the movement as well as the sitting-up activity of the mice are separately determined every fifteen minutes over a period of seventy-five minutes. The E D,50 and E.D.200 are respectivel~, the dosages at which the observed motor activity of the mice is half or twice that of the control animals. Many of the compounds which cause '- 10 - loo-~sal sedation of the mouse can also act as stimulants and the above method permits a~ evaluatlon of the chrono-logical differentiation of the two opposed actlvities on the motor activity o the mous~. .
The compounds of formula I also exhibit anti-depressant activity as indicated in standard -tests in animals for example in the tetrabenazine antagonism test according to G. Stille [Arzneimittelforschung 1~, 534-7 (1964)] in which an antagonism of ptosis and catalepsy induced in rats by tetrabenazine is obsPrved.
From the aforementioned pharmacological tests it can be determinad that the following compounds of formula Ia, A' - B - C' -D - ~ Ia wherein A' is H-Trp, and C' is Gly, Ser; -NH-CH-C0-or -NH-CH-C0-o primarily suppress central nervous system activity whereas the remaining compounds of formula I primarily stimulate the central nervous system.

:

76~3 oo ~88 The compounds which suppress CNS activity are indicated for use as sedatives and especiall~ as soporifics, whereas the compounds whlch stimulate the CNS are indicated for use:Ln the t-eatment of cerebral insufficiency and depress:Lon.
For these uses ~n indicated daily dose is from about 5 to about 500 mg, conveniently adminis-tered in unit dosage form, contalning ~rom about 1.25 to about ~50 mg, or in sustained release form.

The compounds may be adminis~red in p~.armaceutically acceptable salt foxms including acld addition salt forms, or ln the form of complexes.
Such forms exhibit the same order of actlvity as the free base forms and are readily prepared in conven-tional manner. Representative aclds for acid addltlon salt formation lnclude organic aclds, such as trifluoroacetlc acld and lnorganic acids such as hydrochloric acid. Sultable metals for complex foxma-ti`on include calcium and magnesium.

The present invention also provides a pharmaceutical composltion comprislng a compound of formula 1, ln ~ree base form or in the form of a pharmaceu,tically acceptable salt or complex, in association wlth a pharmaceutlcally acceptable carrier ~L187~i8 ~ 100-4~1 or diluent. Such compositions may be in the form of, for example, a solution or capsule.
In one group of compounds A is H-Trp, H-MeTrp, H-Trp~5-OH~ or ~-MeTrp(5-OH)~ B is Ala~
C is Ser, Thr, ~la, Gly or a residue of formula -NH-CH-CO-or -NH-CH-CO-o . whereln R is hydrogen, alkyl of 1 to 4 carbon atomsan alkali metal or an alkaline earth met 1, D is Gly and E is Asp-OH, Asp-OR', Asp-NH~, Asn-OH, Asn-OR', Asn-NH2~ asparaginol or a residue of formula ~ H2 CH2 OH
C}~2oR
wherein R' ls alkyl Of 1 to 4 carbon atoms and the - res~dues A, ~, C and E have the L-, D- or D, L-configuration. - -. , . . . . .. . . . .. . . .. .. . _ . .. _ .
In a second group of compounds A is ~-~rp, H-MeTrp, H-Trp(5-OH) or H-MeTrp, B is Ala, C ls Se~, Thr, Ala, Gly or a residue of formula -NH-CH-CO-or -N~-CH-CO-O

~. ~

8~
~ - 13 - 100~8~31 wherein R is hydrogen, alkyl of 1 ~o 4 carbon atom~, an alkali metal or an alkalln~ earth metal, D ls Gly and E is Asp(OR')-X' wherein R' 1 alkyl o~ 1 to 4 carbon atoms and X' is hydroxyl, alkoxy o~ 1 to 4 carbon atoms or NH2 and the residues A, B, C and ~s~ X' have the L-, D- or D, L- confi~uration.

Claims

THE EMBODIMENTS OF THE INVENTION IN WHICH AN
EXCLUSIVE PRIVILEGE OR PROPFRTY IS CLAIMED ARE
DEFINED AS FOLLOWS:

1. A process for the production of a compound of formula I, A - B - C - D - E I

wherein A is H-Trp, H-MeTrp, H-Trp(5-OH) or H-MeTrp (5-OH), B is Ala, C is Ser, Thr, Ala, Gly, Val, or a residue of formula or wherein R is hydrogen, alkyl of 1 to 4 carbon atoms/ or an alkall metal or an alkaline earth metal and Rl is hydrogen or methyl, D is Gly ox Sar, E is a residue wherein X and Y independently signify COZ or CH2OH, Z is OH, OR2, NH2, NHR3 or NR3R4, and each of R2, R3 and R4 independently signifies alkyl of 1 to 4 carbon atoms, whereby the residues A, B, C and E can have the L-, D- or D, L configuration, as well as the pharmaceutically accept-able salt forms and complexes thereof, which process comprises, a) removing at least one protective group from a protected peptide having the sequence indicated in formula I, or b) linking together by an amide bond two peptide units each of which contains at least one amino acid and which is in protected or unprotected form, the peptide units being such that the amino acid sequence given in formula I is obtained and, if necessary, carrying out process variant a), or c) converting a group E of an unprotected or protected peptide into another group E having the definition pre-viously indicated and, if necessary, effecting process variant a), and, if desired, converting a compound of formula I thus obtained into a pharmaceutically acceptable salt form or complex thereof.

2. A process according to Claim 1 wherein in the compound of formula I A is H-Trp and C is Gly, Ser, or

3. A process according to Claim 1 wherein in the compound of formula I A is H-Trp, H-MeTrp, H-Trp(5-OH) or H-MeTrp(5-OH), B is Ala, C is Ser, Thr, Ala, Gly or a residue of formula or wherein R is hydrogen, alkyl of 1 to 4 carbon atoms, an alkali metal or an alkaline earth metal, D is Gly and E
is Asp-OH, Asp-OR', Asp-NH2, Asn-OH, Asn-OR', Asn-NH2, asparaginol or a residue of formula wherein R' is alkyl of 1 to 4 carbon atoms and the residues A, B, C and E have the L-, D- or D, L-configuration.

4. A process according to Claim 1 wherein in the compound of formula I A is H-Trp, H-MeTrp, H-Trpt(5-OH), or H-MeTrp(5-OH), B is Ala, C is Ser, Thr, Ala, Gly or a residue of formula or wherein R is hydrogen, alkyl of 1 to 4 carbon atoms, an alkali metal or an alkaline earth metal, D is Gly and E is Asp(OR')-X' wherein R' is alkyl of 1 to 4 carbon atoms and X' is hydroxyl, alkoxy of 1 to 4 carbon atoms or NH2 and the residues A, B, C and Asp(OR')-X' have the L-, D- or D, L-configuration.

5. A process according to Claim 1 wherein the com-pound of formula I has the structure H-Trp-Ala-Ser-Gly-Asn-ol or H-Trp-Ala-Ser-Gly-Asp-ol,

6. A compound of formula I as defined in any one of Claims 1 to 3 or a pharmaceutically acceptable salt form or complex thereof, whenever prepared by a process as claimed in any one of Claims 1 to 3 or by an obvious chemical equivalent thereof.

7. A compound of formula I as defined in Claim 4 or 5 or a pharmaceutically acceptable salt form or com-plex thereof, whenever prepared by a process as claimed in Claim 4 or 5 or by an obvious chemical equivalent thereof.